TABLE 1.

Bacterial strains, plasmids, and primers used in this study

Strain, plasmid, or primerRelevant characteristic(s)Source or reference
Strains
    Serratia marcescens
        CH-1Clinical isolate 27
        PC100CH-1 dapASm::Km, KmrThis study
        PC101CH-1 nlpBSm::Sm, SmrThis study
        PC102CH-1 dapASm::Sm, SmrThis study
    Escherichia coli
        CC118 λpirλ-pir lysogen of CC118 [D(ara-leu) araD DlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1]; permissive host for suicide plasmids requiring the Pir protein 22
        S17-1 λpirλ-pir lysogen of S17-1 [thi pro hsdR hsdM+recA RP4 2-Tc::Mu-Km::Tn7 (Tpr Smr)]; permissive host able to transfer suicide plasmids requiring the Pir protein by conjugation to recipient cells 22
Plasmids
    pACYC184Cloning vector, P15A replicon, Tcr, Cmr 22
    pBCSK+Cloning vector, ColE1 replicon, AmprStratagene
    pUT-SmSuicide plasmid requiring the Pir protein for replication and containing a mini-Tn5 cassette containing Smr genes 22
    pBAD18-CmCloning vector, arabinose regulation; CmrColin Hughes (Cambridge, United Kingdom)
    pBG200pBAD18-Cm::dapASm; CmrThis study
    pBG201pBAD18-Cm::nlpBSm; CmrThis study
    pBG203pBAD18-Cm::dapASm nlpBSm; CmrThis study
    pBG401pACYC184 (PshlBASm::luxAB), CmrThis study
    pSC300pUT-Sm derivative carrying nlpBSm::Smr insertion, SmrThis study
    pSC301pUT-Sm derivative carrying dapASm::Smr insertion, SmrThis study
Primers
    Dapk15-ATCTAGACGCGCGAGCCTGAAAA-3
    Dapk25-ATCTAGAGCGAAGGCACGTTATACAGG-3
    NlpBk15-AAGCTTCGCCTGCAGCAGGC-3
    NlpBk25-AAGTCGACGGTAGCAAAAGTAGTG-3
    NlpBk35-GATATCGCTGGTTTTGCTGAAGGC-3
    NlpBk45-AAGCTTGCAGAGCGGCGCCG-3